Objective To investigate the mechanism and role of ?-ray of 103Pd in the treatment of biliary duct cancer.Methods A series of biliary duct cancer cells were treated with different ?-ray dose,and MTT [3-(4,5-dimethy thiazol-2-yl)-2,5-diphenyl terazolium-bromide] technique was used to determine the inhibition rate of ?-ray of 103Pd on the biliary duct cancer cells;and electron micro-technique,DNA agarose gel electrophoresis and flow cytometry to evaluate the morphological characteristics and apoptosis rate of the biliary duct cancer cells were also used.Results The ?-ray radiation of 103Pd resulted in significant inhibition of the biliary duct cancer cells.The features of biliary duct cancer cells apoptosis(e,g:apoptic bodies,DNA ladders band hypodiploid DNA peak) could be seen in the group with lower dosage(5.333mci),and cell necrosis was seen in higher dosage(more than 6.645 mci).Conclusions The ?-ray radiation could induce apoptosis of the biliary duct cancer cells,but with dose dependence,and apoptosis can be an important mechanism for radiation treatment of biliary duct cancer.

AIMTo investigate the preparation of diclofenac sodium pulsatile release pellets (DS-PRP), the release in vitro and the pharmacokinetics of the drug.

METHODSDiclofenac sodium (DS) core pellets prepared by extrusion-spheronization technology were coated in a mini-fluidized bed spray coater with swelling material as the inner coating swelling layer and ethylcellulose aqueous dispersion as the outer coating controlled layer. The effects of formulation and medium on pulsatile release of DS were investigated under release rate test. Pharmacokinetic and bioavailability study in eight human subjects were performed by HPLC method.

RESULTSThe delayed-release time and release rate of DS from DS-PRP were influenced obviously by the swelling material, the concentration of SDS in medium, the coating level of the inner swelling layer and the outer controlled layer. In vitro, the delayed-release time T0.1 was 3.1 h, and the pulsed-release time T0.1-0.2 was 1.2 h. In vivo, the delayed-release time Tlag was 2.8 h, and the bioavailability was (91 +/- 12)%.

CONCLUSIONThe release of drug from DS-PRP was shown to be in pulsed way both in vitro and in vivo.

Adult ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; pharmacokinetics ; Biological Availability ; Cellulose ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; Diclofenac ; administration & dosage ; pharmacokinetics ; Humans ; Hydrogen-Ion Concentration ; Male ; Random Allocation ; Sodium Dodecyl Sulfate ; chemistry

Viral respiratory tract infection is among the leading causes of mortality and morbidity worldwide. Rapid screening methods for multiple detection of a wider range of pathogens become very important for diagnosis of respiratory infection. This article describes conventional detection technologies and several emerging multiplex assays that have potential applications in the diagnosis and monitoring of respiratory viral infections. These techniques include new rapid culture system, multiplex reverse transcription-PCR, real-time reverse transcription PCR, solid and suspension microarrays, mass spectrometry as well as metagenomics methods. The development and application of these techniques will not only improve the ability of rapid detection and control of viral respiratory infection, but play pivotal roles in the rapid characterization of new viral pathogens.
Animals ; Diagnostic Techniques and Procedures ; Humans ; Mass Spectrometry ; methods ; Microarray Analysis ; methods ; Polymerase Chain Reaction ; methods ; Respiratory Tract Infections ; diagnosis ; virology ; Virus Diseases ; diagnosis ; virology ; Viruses ; classification ; genetics ; isolation & purification

Objective To investigate the inhibitory effects ofPolyphyllin D (Paris saponin [PS]Ⅰ) and formosanin C (PS Ⅱ) on human glioblastoma U251 cells and its mechanism.Methods Glioma cell line U251 was cultured in vitro.Different treatments (PSⅠ and PS Ⅱ) were added into the medium cultured human malignant glioma U251 cells; and blank control group was also established.The effects of PSⅠ and PS Ⅱ on proliferation of glioma cells U251 in vitro was examined using methyl thiazolyl tetrazolium (MTT) assay.The nuclear morphology changes of apoptotic cells were detected by Hoechst33258 fluorescent staining.Apoptotic rate was quantified by flow cytometry (FCM) using Annexin-V/PⅠ dual staining.Western blotting was used to evaluate the changes of Fas,Caspase-8 and Caspase-3 proteins in U251 cells.Results PS Ⅰ and PS Ⅱ inhibted U251 cell proliferation with a timeand dose-dependent manner; as compared with that in the blank control group,the proliferation rate in the PSⅠ and PS Ⅱ treatment groups was significantly increased (P＜0.05); PS Ⅰ had stronger inhibited effect than PS Ⅱ.Typical morphological changes of apoptosis were observed in U251 cells of the PSⅠ and PS Ⅱ treatment groups with Hoechst 33258 staining.The early apoptotic rates of the PSⅠ and PS Ⅱ treatment groups were all significantly higher than that of the blank control groups (F=81.434,P=0.000).As compared with those in the blank control group,the expressions ofFas,Caspase-8 and Caspase-3 were significantly increased (P＜0.05).Conclusion PSⅠ and PS Ⅱ inhibite the proliferation ofglioma cells in vitro; they increase the expressions ofFas,Caspase-8 and Caspase-3,and accelerate the cell apoptosis,which might be the mechanism of inhibition.

Objective To study the dosimetry of different arrangements of 125I seeds in one plane.Methods Nine different in-plane arrangements of 9 125I seeds (2.035 × 107 Bq/seed) were simulated according to distance (cm) along x (horizontal)- and y( longitudinal )-axis using the 3-dimensional treatment planning system (TPS) (3D-TPS): x0.5, y0. 5; x0. 5, y1.0; x0. 5, y1.5; x1.0, y1.0; x1.0, y1.5;x1.5, y1.5; x0. 5, y0. 5 (2)1.0; x0.5, y1.0 (2)0.5; x1.0, y1.0 (2)0.5. The isodose curves of 40,80, 130, 145 and 200 Gy were created and the area, radius and medical cost under the 40, 80, 130, 145and 200 Gy isodose curves were calculated. Results The area, radius and medical cost under the same isodose curves were significantly different with each 125I seed arrangement. The arrangements which had the biggest area under curves of 40, 80, 130, 145 and 200 Gy isodose were x1. 5, y1. 5; x1. 0, y1. 0; x1. 0,y1. 0; x0. 5, y1. 0 and x0. 5, y1. 0, respectively. Conclusion The matched peripheral dose and therapeutic effect were affected significantly by the geometric arrangement of 125I seeds.

Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; epidemiology ; virology

The ORF sequence of glycosyltransferase gene BcUGT1 cloned from Bupleurum chinense DC. was analyzed and its three dimentional structure was predicted. Using qRT-PCR method, the expression characteristics of BcUGT1 after methyl jasmonate (MeJA) induction and in different plant tissues were investigated. The results showed that BcUGT1 may be involved in saikosaponin biosynthesis in B. chinense. Thereafter, the recombinant vectors of BcUGT1 were constructed for its expression in E. coli. The target protein was successfully expressed and purified. In the present study, three vectors, pRSET-A, pET-28a (+) and pET-30a (+), and three isolates of E. coli, BL21 (DE3) plysS, BL21A1 and BL21-CodonPlus (DE3)-RIPL were used under different induction conditions, such as different concentrations and during times of inducers (L-arabinose and IPTG) and different inducing temperatures. The results showed that in the condition of 0.5 or 1 mmol x L(-1) IPTG, 16 degrees C, 20 h, target protein expressed in BL21-CodonPlus (DE3)-RIPL with pET-28a (+) or pET-30a (+) as vector. Using PrepEase His-tagged protein purification kit, the target protein was purified. The present work will be helpful for follow-up bio-function analysis of BcUGT1.
Amino Acid Sequence ; Base Sequence ; Bupleurum ; chemistry ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Glycosyltransferases ; genetics ; isolation & purification ; metabolism ; Oleanolic Acid ; analogs & derivatives ; biosynthesis ; Open Reading Frames ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; Protein Structure, Secondary ; Recombinant Fusion Proteins ; genetics ; metabolism ; Saponins ; biosynthesis

Global Health ; History, 20th Century ; Humans ; Influenza A Virus, H2N2 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; transmission ; virology

OBJECTIVETo investigate the role of a potential diabetes-related mitochondrial region, which includes two previously reported mutations, 3243A-->G and 3316G-->A, in Chinese patients with adult-onset type 2 diabetes.

METHODSA total of 277 patients and 241 normal subjects were recruited for the study. Mitochondrial nt 3116 - 3353, which spans the 16S rRNA, tRNA(leu(UUR)) and the NADH dehydrogenase 1 gene, were detected using polymerase chain reaction (PCR), direct DNA sequencing, PCR-restriction fragment length polymorphism and allele-specific PCR. Variants were analyzed by two-tailed Fisher exact test. The function of the variants in 16S rRNA were predicted for minimal free energy secondary structures by RNA folding software mfold version 3.

RESULTSFour homoplasmic nucleotide substitutions were observed, 3200T-->C, 3206C-->T, 3290T-->C and 3316G-->A. Only the 3200T-->C mutation is present in the diabetic population and absent in the control population. No statistically significant associations were found between the other three variants and type 2 diabetes. The 3200T-->C and 3206C-->T nucleotide substitutions located in 16S rRNA are novel variants. The 3200T-->C caused a great alteration in the minimal free energy secondary structure model while the 3206C-->T altered normal 16S rRNA structure little.

CONCLUSIONSThe results suggest that the 3200T-->C mutation is linked to the development of type 2 diabetes, but that the other observed mutations are neutral. In contrast to the Japanese studies, the 3316G-->A does not appear to be related to type 2 diabetes.

Age of Onset ; Aged ; Alleles ; Base Sequence ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Humans ; Middle Aged ; Models, Molecular ; Nucleic Acid Conformation ; Point Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal, 16S ; chemistry ; genetics

OBJECTIVETo develop a rapid duplex Real-time reverse transcription PCR (rRT-PCR) method to detect E119V mutation on neuraminidase (NA) of influenza A(H3N2) subtype with drug resistance to oseltamivir.

METHODSTwenty-six NA genes of influenza A(H3N2) virus between 2000 and 2012 in GenBank database were selected as the target genes, and specific TaqMan-MGB probe was designed to target the E119V amino acid change in neuraminidase protein. rRT-PCR was then performed and evaluated for the sensitivity, specificity and reproducibility using virus with E119V mutation and clinical samples.

RESULTSThis study described the validation of a highly sensitive and specific duplex rRT-PCR for detection of substitutions leading to the E119V amino acid change in NA protein of influenza A(H3N2). Fluorescence signals could be detected even when diluted a A (H3N2) virus (HA = 8) into 10(-5) and linear correlation between the logarithm of the viral titer with the Ct values was observed. In addition, the assay was highly specific in that there was no cross-react with other respiratory viruses, nor did two TaqMan-MGB probes. E119V substitution in quasispecies with both sensitive and resistant viruses could be detected as well. The limit of detection was 5% for quasispecies with high concentrations and 50% for quasispecies with low concentrations. The average coefficient of variation (CV) for within-run assays was 2.32% and 0.57% for H3N2-119E and H3N2-119V primer/probe sets separately, 1.77% and 0.97% for average CV of between-run assays, which exhibited good repeatability. Sequence analysis of twenty NA genes verified glutamic acid (E) at amino acid site 119, which was in consistent with the results from our rRT-PCR method.

CONCLUSIONThe assay developed in this study is highly sensitive and specific, and easy to operate; thereby it could be used for identification of A(H3N2) virus with E119V amino acid change in NA protein.

Amino Acid Substitution ; Drug Resistance, Viral ; Influenza A Virus, H3N2 Subtype ; drug effects ; enzymology ; genetics ; Mutation ; Neuraminidase ; genetics ; Nucleic Acid Probes ; Reverse Transcriptase Polymerase Chain Reaction ; methods